Difference between revisions of "Part:BBa K2967013"

Latest revision as of 09:15, 16 October 2019

Amplifier system with hIL-10.

Amplifier system is created by our team NEU-CHINA 2019. To make sure that amplifier is able to strengthen the expression of gene downstream to promoter hrpL. A coomassie brilliant blue was demonstrated that more protein at 35kDA, which should be IL-10, was witnessed when introducing IL-10 gene downstream of promoter hrpL. This amplifier system tended to strengthen overall protein expression including the human IL-10.

Figure 1. Diagram of expressing vector. HrpR and HrpS amplifier system is used as a booster to boost the expression level of gene downstream promoter hrpL. Protein HrpR and HrpS can bind together to induce the PhrpL which strengthens the expression of downstream gene YebF-IL10.

Figure 2. Protein expression of human IL-10 gene which transformed in BL21 strain. CFU was then inoculated to LB medium followed by 12h incubation at 37℃. After induction of IPTG with the final concentration at 1mM, the culture was incubated at 37℃ for 2h. A coomassie brilliant blue was demonstrated to show the result. The red box shows where the target protein is.

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
COMPATIBLE WITH RFC[12]
21
INCOMPATIBLE WITH RFC[21]
Illegal BglII site found at 2082
23
COMPATIBLE WITH RFC[23]
25
COMPATIBLE WITH RFC[25]
1000
INCOMPATIBLE WITH RFC[1000]
Illegal BsaI.rc site found at 1292
Illegal SapI.rc site found at 1925

@@ Line 3: / Line 3: @@
 <partinfo>BBa_K2967013 short</partinfo>
-Amplifier system is created by our team NEU-CHINA 2019. To make sure that amplifier is able to strengthen the expression of gene downstream to promoter hrpL. A coomassie brilliant blue is demonstrated to show that more protein at 35kDA, which should be IL-10, is witnessed when introducing IL-10 gene downstream of promoter hrpL. And it can be seen that lanes with amplifier  seems to be darker, which means that this amplifier system tends to strengthen overall protein expression.
+Amplifier system is created by our team NEU-CHINA 2019. To make sure that amplifier is able to strengthen the expression of gene downstream to promoter hrpL. A coomassie brilliant blue was demonstrated that more protein at 35kDA, which should be IL-10, was witnessed when introducing IL-10 gene downstream of promoter hrpL. This amplifier system tended to strengthen overall protein expression including the human IL-10.
 https://2019.igem.org/wiki/images/d/df/T--NEU_China--part--amp_passway_use_this.png
-'''Figure 1. Diagram of expressing vector.''' HrpR and HrpS amplifier system is used as a booster to boost the expression level of gene downstream promoter hrpL. Protein HrpR and HrpS can bind together to induce the PhrpL which strengthen the expression of downstream gene YebF-IL10.
+'''Figure 1. Diagram of expressing vector.''' HrpR and HrpS amplifier system is used as a booster to boost the expression level of gene downstream promoter hrpL. Protein HrpR and HrpS can bind together to induce the PhrpL which strengthens the expression of downstream gene YebF-IL10.
 https://static.igem.org/mediawiki/parts/thumb/1/1b/T--NEU_China--part--amp%2Bil-10-1.png/800px-T--NEU_China--part--amp%2Bil-10-1.png
-'''Figure 2. Expressing vector is transformed into BL21 strain. CFU is then inoculated to LB medium followed by 12h incubation at 37℃.''' After induction of IPTG, final concentration at 1mM, the culture is incubated at 37℃ for 2h. A coomassie brilliant blue is demonstrated to show the result. The red box shows where the target protein is.
+'''Figure 2. Protein expression of human IL-10 gene which transformed in BL21 strain. CFU was then inoculated to LB medium followed by 12h incubation at 37℃.''' After induction of IPTG with the final concentration at 1mM, the culture was incubated at 37℃ for 2h. A coomassie brilliant blue was demonstrated to show the result. The red box shows where the target protein is.
 <!-- Add more about the biology of this part here